Histopathological examination of tissue has been an essential component of pathology. Current knowledge about carcinogenesis, cancer diagnostics and prognostication is predominantly based on histological study on tissue, cells and nuclei. Morphometric information, especially nuclear morphometry, has been shown to have great potential in assisting cancer screening, diagnosis, grading and classification, prognosis, analysis of angiogenesis, and evaluating the efficacy of therapy.
Traditional histopathological examination has several limitations, including: (1) it can only acquire the two dimensional information from the cells or nuclei of interest; (2) it is qualitative or semi-quantitative; (3) it requires tissue excision and processing for examination; and (4) it is frequently subject to the inter- and intra-personal variability in interpretation, thus with a relatively low reproducibility. Digital morphometric techniques were introduced to overcome these problems. Digital morphometry helps improve the reproducibility of histological examination, but its clinical utility is yet to be proven. Better segmentation (the delimitation of boundaries between two compartments) and comprehensive examination of the entire cell has been found to be crucial for yielding reproducible and convincing results. Three-dimensional (3D) stereological methods have recently been introduced in clinical diagnostics and are proving to be beneficial.
The majority of human cancers (nearly 90%) arise from the epithelial cells that line many organs. In dysplastic epithelial tissue, dysplastic epithelial cells differ from normal ones in their shapes and the size of their nuclei. A variety of optical spectroscopic and imaging methods can be used to detect abnormal changes in light scattering and absorption properties of tissue to detect carcinogenesis. However, to detect cancer in its earliest stage (precancer) it is crucial to depth-selectively probe the specific areas that are initially involved in neoplastic transformations (e.g. the base of the crypt for colon carcinogenesis). It therefore is highly desirable to reliably and accurately image epithelial cells and their nuclei at various depths.
Current approaches such as diffuse optical tomography (DOT) suffer from poor spatial resolution (5-10 millimeters) due to light diffusion. Other approaches such as Optical Coherence Tomography (OCT) have yielded micrometer resolution and cross-sectional imaging. This permits the imaging of tissue microstructure in situ, yielding micron-scale resolution image with use of low temporal coherence light. However, it is difficult for OCT to image structures such as nuclei for various technical reasons.
It is desirable to combine the advantages of both techniques: (1) the sensitivity to nuclear morphology and cellular structure of light scattering, and (2) the high spatial resolution offered by low coherence light, to perform high resolution imaging of the structure and composition of tissue. This would provide a 3D image of the nuclear morphology and cellular structure for tissue in real time, with no tissue excision or processing required.